METHOD FOR PRODUCING NUCLEAR proteinases from non-histone proteins and histone PLANTS

摘要

A method for producing nuclear proteinases from non-histone and histone proteins of plants, comprising separating at specified time intervals from the start of soaking 0 h, 3 h, 6 h, 9 h, 12 h, 15 h, 18 h, 21 h embryos from the endosperm followed by preservation embryos buffered in 80-90% glycerol at -25 ° C, separation of cell nuclei, nuclear extraction suprastructures increasing concentrations of 0.14 M, 0.35 M, 2 M sodium chloride and 6 M guanidine hydrochloride with 0,1% β-mercaptoethanol, selection of these fractions and non-histone proteins with histone n omoschyu ion exchange chromatography with Amberlite IRC-50 in a discontinuous gradient of guanidine hydrochloride 6%, 8.9%, 10.6%, 13%, 40% 0.1 M potassium phosphate buffer, pH 6.8, with isolation of nuclear proteinases affinity chromatography using the method through the column with sepharose 4B with immobilized trypsin inhibitor followed by evaluation of proteolytic activity.